LOCUS       sequence-374283-        9461 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Backbone vector for generation of BiFC plasmids.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9461)
  AUTHORS   Subotic A, Swinnen E, Demuyser L, De Keersmaecker H, Mizuno H, 
            Tournu H, Van Dijck P
  TITLE     A Bimolecular Fluorescence Complementation Tool for Identification 
            of Protein-Protein Interactions in Candida albicans.
  JOURNAL   G3 (Bethesda). 2017 Oct 5;7(10):3509-3520. doi: 
            10.1534/g3.117.300149.
  PUBMED    28860184
REFERENCE   2  (bases 1 to 9461)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1534/g3.117"; 
            journalName: "G3 (Bethesda)"; date: "2017-10-5- 5"; volume: "7"; 
            issue: "10"; pages: "3509-3520"
COMMENT     Sequence Label: 2016-C
FEATURES             Location/Qualifiers
     source          1..9461
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     90..107
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    224..245
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(260..265,266..283,284..290)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    298..314
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     303..325
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     322..338
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     322..338
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        356..374
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     356..373
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             4742..5044
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     4923..4942
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             5393..6187
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             5393..6187
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5447..5466)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6057..6076
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     regulatory      6387..6396
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     rep_origin      8809..9397
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9298..9317
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
       61 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
      121 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
      181 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
      241 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
      301 tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc aagctattta
      361 ggtgacacta tagaatactc aagctatgca tcaagcttgg taccgagctc ggatccacta
      421 gtaacggccg ccagtgtgct ggaattcgcc cttaagaaca tttctgtacc gcaaatgtat
      481 cgactttttc gatggtgtgt tattgttgtt ttgttgcgaa atataatagc ttctgcttta
      541 tatcctttgt gttgatcctt cccctccttt catacttgta aaattacaat tggtattttc
      601 aaccaatatt accacacgtc atgcactaac tcaaatgggc aaaaaaaaga gagataggaa
      661 ttacgtcatt ttattatgtt tttacgtgga ctagaaggtt aagataattt ctggttattt
      721 tgttgataat tgagttttgt tcgacctcca ccaatcttta aactctcgtt gagtattcat
      781 tttaatgatt ttttgaaatc agaaggggtg tttctaaccc aggattgttt tattgagagg
      841 ttgaaacatc aatgtgttag tttattgatt tgggttggtt taatagaaaa atatcagtag
      901 gtgttcaata attgtagttt attttcttgg ctaaaattgt tgcgaaaaat tgacaaaata
      961 ataacaaaaa gtattttaga tgctaaaaca taatagtagg aatatctacc tgaaccattt
     1021 acattcaaca actggcaaga accttcctaa taatttgcaa gagtaatcac aattaaacca
     1081 aaatcaaagt ttaccgtgta tatagttgga cagaaagctg tcacattggt ttaatacata
     1141 gttgaatatg tatgcatgaa acaacaacaa tacaattaca gctcatgatc cttcaattaa
     1201 tttcaacatt caaacaacat aacaccttgt gtaaatgagg ttatgagtac atgtgaacaa
     1261 caaatcttat gactaaactc gggtcgtgta cttttgacga ttgagtcatt tttaccaatt
     1321 aggcaattaa tttttgctaa attttcatta accgcgctgg ccatcccaac aaacaatata
     1381 aaagctgaca acgaaaaaaa aaaaaaaaaa aactccaatt ctgaaaaatt tttcttcttc
     1441 ttcatactaa acaaaacaac tataaagata atcaattaat aatgtcacaa caacctagtg
     1501 aaaataaatt atggggtggt cgattcactg gcgccactga tccattaatg gatttataca
     1561 atgcttcatt accttatgat aaagttatgt atgatgcaga tttaactggt actaaagtat
     1621 atactcaagg attgaacaaa ttaggattaa ttaccactga agaattgaat ctgattcatc
     1681 aaggtttaga acaaattaga caagaatggc atgataataa atttattatt aaagctggtg
     1741 atgaagatat tcatactgct aatgaacgta gattaggaga aatcattggg aaaaacattt
     1801 ctggtaaagt tcatactggt agatccagaa atgatcaagt tgctactgat atgagaattt
     1861 ttgttcgtga atcattactt aatttaagta agattttaca tcaattcatt acagcaattc
     1921 ttgaacgagc tcataaagaa attgatgtat taatgcctgg ttatactcat ttacaaaaag
     1981 ctcaacctat tagatgggct cattggttaa gttcatatgc aacttatttc actgaagatt
     2041 ataaacgttt acaagaaatc attactcgtg ttaatcaatc tccattagga tctggtgctt
     2101 tagctggtca tccctacggt attgatcgtg aatttttagc taaaggatta ggatttgatg
     2161 gtgttattgg gaattcttta actgcagttt ctgatcgtga ttttgttgtt gaaagtttat
     2221 tttggagtac attatttatg aatcatattt ctcgattttc agaagattta attatttatt
     2281 ctagtggtga atttggattt ataaagttag ctgatgctta ctctactggt tcatcattaa
     2341 tgccacaaaa gaaaaatcct gattcattag aattattacg tgggaaaagt ggcagagtgt
     2401 ttggtcagtt atctgggttt ttaatgtcaa ttaaatccat tccatcaacc tataataaag
     2461 atatgcaaga agataaagaa ccattatttg atgcattgac taccgtagaa cattcaattt
     2521 taattgctac tggtgtgatt tctactttac tgattgataa acaaaatatg gaaaaagctt
     2581 taactatgga tatgttagct actgatttag cagattattt agttagaaaa ggtgttccat
     2641 ttagagaaac tcatcatatt tctggtgaat gtgttagaaa agctgaagaa gaaaaattgt
     2701 cgggaattga ccaattatca tttgaacaat ttcaacaaat tgattcacgt tttgaaaaag
     2761 atgttatgga aacttttgat tttgaagcta gtgtggaaag aagagatgct attggtggta
     2821 ctgctaagag cgctgtttta aaacagttag aaaatttgaa atcaatttta aattaattta
     2881 taaataatca tatagtattc aatgttttgt tttgtttttt tttttttttt natggaaatg
     2941 tacaaacagt atataatcaa ttaatagaaa gataccctgt attccattgt atcgcctttg
     3001 ggatgtctat aataccaact ttcaagaatg agaaattaaa caacaatttc tttcaccaat
     3061 caactcgtct atattcaata aactcattct ggtcttttca aatatttaat gttatcctga
     3121 tcaaacttac aaatcaaaaa tgtatcaaaa tcattcatta gttcttcttc ttttgttatc
     3181 tttgtcacta tatttctctc catccagacc aaaaatactt ccattgtgaa gatcgtctga
     3241 atcataatca gcaactacct cttccatttc agagaattca gatttttcaa aataggtatc
     3301 acgggccaag tacacacaag acaaacgata cgagtgctgg gatatttctt tcaatgtatc
     3361 actaaattcg ttgagattta aagagagccc tttgactgct ttcattactt tcctcacatc
     3421 atgagcatcc acattcaaat ttcctgtagc cctttattct tagttttctt ttcttattct
     3481 tttcttcgta cattgaaatc acaaatggtg atatcaaacg aaggtcacac tgacttatgt
     3541 ctaatacctc caatgatgat actggtaact taaccaggtc tactcttttc aaacgatttc
     3601 tcattagtgt caatttacgt aggttcttca acctttcaaa cgatgcaaat agcttttgta
     3661 agggcgaatt ctgcagatat ccatcacact ggcggccggc cgctcgagca tgcatctaga
     3721 gggcccttga agaattggaa gaagttagta gttaaaaaaa caatgggaaa gatttaatct
     3781 ttttttcttt tttcaaaaat ttttaaccgc tgatgatata ttaatctctc tccaccactc
     3841 tctttaaaca ccacaattat taataataag atatctctcc tactagttat ttgttagtaa
     3901 gtatattgtt gattgattgg ttgggttaaa aatttttttt ttcataaaga aaataatgaa
     3961 agtgagaaaa atttaaaccc cacattacat actcaacaca acactacaca cacgcattta
     4021 acaatcatca tcagttgaaa tatattttac aaacaattac gattgtttaa tgaaaactat
     4081 ttctgttgtt attattattt aacctttttt ttcttgttgg gttgaccctt tttctattaa
     4141 ctatctaact accacataac atttgaagtt gaactattaa accttaattt gcggccgcaa
     4201 atctaatcta ctttcttctt cttctaccct atttattctg aaatgggggg gggggggggt
     4261 ggtgggaact ttaggcgtgt taaatttgtc acatctgttc tcacttgctt ctattcgtat
     4321 aattttaatt gtcttgtgag atcaatcttg ttcttgttcg agttttttct tgctttgatt
     4381 aactactatt attattagta tgcaatgctg tttattacca tttcataact tatattataa
     4441 ctcattgatt tcaatacctg ctcaacaatg agtcgttatt attactgtgc tctgtattct
     4501 gtactgctgc acgtacgtta tgtccattga agacaagcaa agaaagacca acattcaatt
     4561 gtatctgctt accagagttg tttacctctg ttaccaatat ttggaaatta attttttggt
     4621 ggtggacgtt tcttcctaaa acacctagat gtgctagtat aattgcacaa ccagcacgca
     4681 cactgcgtct ggccggtcct ggactacatt ttgtctctaa cattcgtagc tgcacgtacg
     4741 gcagtttaag gtttacacct ataaaagaga gagccgttat cgtctgtttg tggatgtaca
     4801 gagtgatatt attgacacgc cggggcgacg gatggtgatc cccctggcca gtgcacgtct
     4861 gctgtcagat aaagtctccc gtgaacttta cccggtggtg catatcgggg atgaaagctg
     4921 gcgcatgatg accaccgata tggccagtgt gccggtctcc gttatcgggg aagaagtggc
     4981 tgatctcagc caccgcgaaa atgacatcaa aaacgccatt aacctgatgt tctggggaat
     5041 ataaatgtca ggcatgagat tatcaaaaag gatcttcacc tagatccttt tcacgtagaa
     5101 agccagtccg cagaaacggt gctgaccccg gatgaatgtc agctactggg ctatctggac
     5161 aagggaaaac gcaagcgcaa agagaaagca ggtagcttgc agtgggctta catggcgata
     5221 gctagactgg gcggttttat ggacagcaag cgaaccggaa ttgccagctg gggcgccctc
     5281 tggtaaggtt gggaagccct gcaaagtaaa ctggatggct ttcttgccgc caaggatctg
     5341 atggcgcagg ggatcaagct ctgatcaaga gacaggatga ggatcgtttc gcatgattga
     5401 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     5461 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     5521 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga
     5581 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     5641 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     5701 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     5761 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     5821 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     5881 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga
     5941 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     6001 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     6061 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     6121 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     6181 cttctgaatt attaacgctt acaatttcct gatgcggtat tttctcctta cgcatctgtg
     6241 cggtatttca caccgcatca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     6301 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     6361 tgcttcaata atagcacgtg aggagggcca ccatggccaa gttgaccagt gccgttccgg
     6421 tgctcaccgc gcgcgacgtc gtctattcca agcctgtggc tttttttggt gattcacaat
     6481 acataaattg gcaaatacct cttctataga ctttaacctt tgatgaaatg gtatggataa
     6541 accacttttg tggtcagggt atatatattt ttattccaat ctggccttta taaattcaaa
     6601 tttccaaggg gactctgcta aaccttctga tataactata actctaataa tggctgtaaa
     6661 aagtgtgaaa aaaagatgat attcacaaaa aagataaagg ggatttcata ttgggtatct
     6721 agatttaaac atcgtatttc aacaatgaaa ctagaatcaa tacgaaatcc agggcacaac
     6781 tgtgttcatt cccttgggaa ttatactgag taatgggaag aatgtgcaaa aaatacccgt
     6841 gtcttatata atttccattt gtgtagggct taaaagtgga caacttcagg ttacgaaacc
     6901 acgaaaatgt ctaaacgttt gatcaaattc gtaactcatg gaaaatcaca gcaacaaatt
     6961 aaggatggat atatccggta cttacaaata aaaacactaa agaagtataa attttgaact
     7021 gaaattctat agaaaatctt tttttttggg taaaattatg cgattgtggc tcatagtaac
     7081 gtaatcatac agttcaacaa acatcatttt gtcaaaggat ttctaatcct attctttttc
     7141 tacttggctt ttgagtatat atatatatat atatacatcc tttgttcatc atcatgttcg
     7201 attgaatttt tttcttggtt gttattagat tgtctacttg agtttttctt ggtgcaaatt
     7261 gtggtgtttt tctaacaaat ttttcaacat ttaaagtatt tctggaactt ttctacaagg
     7321 aacgacatag atccattaat gcgcccctct aaaacaatac ccataatgtc ttcttaagta
     7381 tgcatctgct tataaatgtt tctggattta ctttagttag aatgtcgaca cgacaatgaa
     7441 ctattgctgt agatcacgtg cgatcactga attatgagtt tctttttatt tttctttttg
     7501 ggcttttttt tttgcaattt tttttttcca cctttgccaa ttgcgtagtc acatataatg
     7561 tatacaattt ttgcaactct gacactataa ttgatccaat aataaagtat ataatagatc
     7621 atcaagtata cgtaatctcc ccaaaaaaaa aattgcataa aataaaatct ataaaccttt
     7681 tttagatttt tagttgttcc ttttttcttt ctattcttca acttccaagt gttgtcactt
     7741 tcttcctgtt aactatattt aaaagtaaat accctcccca gaaaactccg gaatgagcct
     7801 agggacccgg gggcgcgccg gaggtggagg ttctggtgga ggtggttcag ctagcacgcg
     7861 tagtacttaa gatcctgaag acgggatcgt tcgaagagtg aaattctgga aatctggaaa
     7921 tctggttttg tattcttgtt attcttcttt ttgttattac atatataact tgttactttt
     7981 ttaaaaaaat ctttgtttat tttataaata tataaaacta aatttaagaa aaagagaaaa
     8041 atgttttatt tgagagattg aaattttact tgaatttagc ttagctttta taaagtatta
     8101 ttatgtaaaa aaacaaaaca aatatacatt aaaaagttaa gactataaaa tagccaccca
     8161 aggcatttct atatcttgtt gttgttgttt tcatcttctg tatcagagga acttatttta
     8221 ttattttcgt cacgggtatt ttctcttgtt tgatgattca tcccattcat tccatcataa
     8281 aattctagaa ggaccacctt tgattgtaaa tagtaataat taccaccctt atttaattat
     8341 ttatttaact tatttattta tttattatac atatatacaa atctaataaa gtgaaaatct
     8401 cccccttcac accggtgtgg tccgggacga cgtgaccctg ttcatcagcg cggtccagga
     8461 ccaggtggtg ccggacaaca ccctggcctg ggtgtgggtg cgcggcctgg acgagctgta
     8521 cgccgagtgg tcggaggtcg tgtccacgaa cttccgggac gcctccgggc cggccatgac
     8581 cgagatcggc gagcagccgt gggggcggga gttcgccctg cgcgacccgg ccggcaactg
     8641 cgtgcacttc gtggccgagg agcaggactg acacgtgcta aaacttcatt tttaatttaa
     8701 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     8761 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     8821 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     8881 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     8941 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     9001 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     9061 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     9121 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     9181 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     9241 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     9301 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     9361 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     9421 acggttcctg gccttttgct ggccttttgc tcacatgttc t
//
